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¹ Department of Cell Biology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada
² Instituto de Investigaciones en Biologiá Experimental, University of Guanajuato, Guanajuato, Mexico
³ The Institute for Systems Biology, Seattle, WA 98103

Address correspondence to Richard A. Rachubinski, Department of Cell Biology, University of Alberta, Medical Sciences Building 5-14, Edmonton, Alberta T6G 2H7, Canada. Tel: (780) 492-9868. Fax: (780) 492-9278. E-mail: rick.rachubinski{at}ualberta.ca

The peroxin Pex24p of the yeast Yarrowia lipolytica exhibits high sequence similarity to two hypothetical proteins, Yhr150p and Ydr479p, encoded by the Saccharomyces cerevisiae genome. Like YlPex24p, both Yhr150p and Ydr479p have been shown to be integral to the peroxisomal membrane, but unlike YlPex24p, their levels of synthesis are not increased upon a shift of cells from glucose- to oleic acid–containing medium. Peroxisomes of cells deleted for either or both of the YHR150w and YDR479c genes are increased in number, exhibit extensive clustering, are smaller in area than peroxisomes of wild-type cells, and often exhibit membrane thickening between adjacent peroxisomes in a cluster. Peroxisomes isolated from cells deleted for both genes have a decreased buoyant density compared with peroxisomes isolated from wild-type cells and still exhibit clustering and peroxisomal membrane thickening. Overexpression of the genes PEX25 or VPS1, but not the gene PEX11, restored the wild-type phenotype to cells deleted for one or both of the YHR150w and YDR479c genes. Together, our data suggest a role for Yhr150p and Ydr479p, together with Pex25p and Vps1p, in regulating peroxisome number, size, and distribution in S. cerevisiae. Because of their role in peroxisome dynamics, YHR150w and YDR479c have been designated as PEX28 and PEX29, respectively, and their encoded peroxins as Pex28p and Pex29p.

Key Words: biogenesis; peroxin; protein similarity; open reading frame; membrane fission

^* Abbreviations used in this paper: DsRed, Discosoma sp. red fluorescent protein; PBD, peroxisome biogenesis disorder; PNS, postnuclear supernatant; PTS1, peroxisome-targeting signal 1; SM, synthetic minimal.

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